Summary. Micellar liquid chromatography (MLC) is a branch of high – performance liquid chromatography. The applications of MLC for the determination of numerous compounds in pharmaceutical formulation, biological samples, food, enironmental samples and feeds have been growing very rapidly. MLC technique has several advantages over other chromatographic techniques. Its main advantage is the small amount of organic modifiers used such as acetonitrile and methanol and the safty and recyclabity of the mobile phase. In this article presents application method with the use of micellar liquid chromatography for analysis an antibacterial substances in biological matrices such as milk, eggs, tissues, and feed.